MMP-8 (also known as type neutrophil collagenase or collagenase 2) has substrate specificity for native  collagens (types I, II and III). MMP-8 was originally found in polymorphonuclear leukocytes (PMN, neutrophils), present in storage granules and is only secreted upon activation of these cells. More recently it was discovered that MMP-8 is also produced by other cells, such as endothelial cells, chondrocytes and synovial fibroblasts. Human MMP-8 is produced and secreted as an inactive precursor form. Neutrophil pro-MMP-8 has a Mw of 75 kDa, whereas other cells may produce smaller 50-55 kD forms of pro-MMP-8, the differences originate by variations in glycosylation.  The activity of MMP-8 is dependent on Zn2+ and Ca2+.  ProMMP-8, can be activated in vitro by organo mercurial compounds such as p-aminophenyl mercuric acetate (APMA), oxidizing agents like hydrogen peroxide or hypochlorite and proteases such as trypsin, chymotrypsin, cathepsin G, tissue kallikrein and MMP-3.   TIMP-1 or TIMP-2 inhibit MMP-8 activity by binding in a 1 to 1 molar ratio.

Studies support use of MMP-8 as a biomarker in periodontal disease, sepsis, and cancer.

The QuickZyme human MMP-8 activity assay enables you to specifically measure in biological samples both active human MMP-8, as well as (pro)MMP-8 which is activated on the plate by APMA.  It can be used for the measurement of MMP-8 activity in various biological samples, such as conditioned culture media, tissue homogenates, serum, plasma and urine.

This product ships on dry ice.


Assay Principle

Active MMP-8    Measurement of active human MMP-8 Standards, controls and biological samples are pipetted into the pre-coated plate. Human MMP-8 present in the biological sample is captured by the antibody. After washing, the pro-detection enzyme is added. This is activated by the active MMP-8 into an active detection enzyme. The active detection enzyme is able to cleave the chromogenic substrate, resulting in generation of a yellow color that can be measured at 405 nm using an ELISA plate reader.

Total human MMP-8       Measurement of total MMP-8 is done similarly to the measurement of active MMP-8.  After binding of MMP-8 to the antibody-coated plate, bound MMP-8 is first activated by adding APMA, resulting in the activation of pro-MMP-8. The activity of total MMP-8 (the newly activated MMP-8 and the already active MMP-8 present in the sample) is measured by adding the detection enzyme, followed by the addition of chromogenic substrate. The released color can be measured at 405 nm using an ELISA plate reader.

Each 96 well kit contains:

  • 96 well microwell plate – 12×8 well ready-to-use strips coated with F(ab’)2 goat anti-mouse
  • Mouse-anti-MMP-8 – 60 µl of 20 µg/ml in assay buffer
  • Assay buffer – 125 ml bottle contains 100 ml ready-to-use TrisHCl buffer
  • Standard – tube contains 50 µl of 480 ng/ml pro-MMP-8 (human)
  • p-Aminophenylmercuric acetate (APMA) – tube contains 17.5 mg APMA
  • Detection enzyme – tube contains 600 µl detection enzyme in Tris-HCl buffer
  • Substrate – tube contains 1000 µl peptide substrate in demineralized water
  • Wash buffer – 30 ml bottle contains 25 ml 20x concentrated phosphate buffer


Other materials required but not supplied:

  • Single and/or multichannel pipettes with disposable polypropylene tips.
  • Polypropylene tubes (Eppendorf tubes).
  • Glass measuring cylinder 500 ml.
  • Distilled or demineralized water.
  • Microplate shaker.
  • Refrigerator at 2-8 °C.
  • Dimethyl Sulphoxide (DMSO).
  • (Microtitre plate) incubator at 37°C.
  • Automatic plate washer or wash bottle (optional).
  • Microplate reader capable of measuring at 405 nm.
  • Measures endogenous active MMP-8 (naturally occurring) or total active MMP-8 (following activation with APMA).
  • Samples: cell culture conditioned medium, serum, plasma, urine, saliva and tissue homogenates.
  • Quantitative.
  • Range: 10 – 24000 pg/ml.
  • Sensitivity: 100 pg/ml for 2 h incubation with detection reagent; 24 pg/ml for 6 h incubation with detection reagent; 4 pg/ml for 24 h incubation with detection reagent.
  • Ease-of-use: Equivalent to ELISA.
  • Marker for neutrophils (PMN)
  • Store kit at -20°C, store standard at -70°C